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Fig. 4 | Biotechnology for Biofuels

Fig. 4

From: Acyl-lipid desaturases and Vipp1 cooperate in cyanobacteria to produce novel omega-3 PUFA-containing glycolipids

Fig. 4

Lipid molecular species by LC–MS/MS of PUFA distribution in engineered and wild-type Leptolyngbya BL0902. LC–MS/MS was conducted on lipid extracts dissolved in isopropyl alcohol/methanol (50:50), chromatographed on an Accucore C30 column, and introduced by heated electrospray ionization into a Q Exactive HF Hybrid Quadrupole–Orbitrap Mass Spectrometer with MS scans collected in data-dependent mode, as described in Methods. a Venn diagram of the number of distinct molecular species observed for Leptolyngbya BL0902 without (WT) or with conjugation with pDBV (n = 4 for each). b, c 18- and 20-carbon PUFAs were observed to be complexed to monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), or phosphatidylglycerol (PG). (B) Distribution of ALA (18:3, n-3), SDA (18:4, n-3) and ETA (20:4, n-3) among the three glycolipids is shown, illustrating the selectivity for the lipid backbone for each PUFA. c Shown in light blue (WT) and dark blue (engineered with pDBV) are the mean ± standard deviation of estimates of mg/g of fatty acids based on (i) normalized peak areas from LC/MS, (ii) fraction of total peak area for each species in a sample, and (iii) known total fatty acid yield for that organism from GC–FID analysis. This treatment assumes that all species exhibit the same ionization efficiency. Species across the bottom refer to the two acyl chains associated with MGDG (M), DGDG (D) or PG. Note the shift from fewer to more double bonds upon introduction of the pDBV plasmid

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